Folin-Ciocalteu Method Research Articles

Phenolic Characterization of a Purple Maize (Zea mays cv. "Moragro") by HPLC-QTOF-MS and Study of Its Bioaccessibility Using a Simulated In Vitro Digestion/Caco-2 Culture Model.

The present work aimed to characterize the phenolic and antioxidant content of the Argentinian purple maize "Moragro" cultivar. Additionally, the INFOGEST simulated in vitro digestion model was used to establish the effect of digestion on bioactive compounds. Finally, digestion samples were used to treat Caco-2 cells in the transwell model to better understand their bioavailability. Twenty-six phenolic compounds were found in purple maize cv. "Moragro", 15 nonanthocyanins and 11 anthocyanins. Several compounds were identified in maize for the first time, such as pyrogallol, citric acid, gallic acid, kaempferol 3-(6″-ferulylglucoside), and kaempferol 3-glucuronide. Anthocyanins accounted for 24.9% of total polyphenols, with the predominant anthocyanin being cyanidin-3-(6″ malonylglucoside). Catechin-(4,8)-cyanidin-3,5-diglucoside and catechin-(4,8)-cyanidin-3-malonylglucoside-5-glucoside were detected as characteristics of this American maize variety. Total polyphenol content (TPC; by the Folin-Ciocalteu method), HPLC-DAD/MSMS, and antioxidant activity [by DPPH and ferric-reducing antioxidant power (FRAP)] were evaluated throughout in vitro digestion. TPC, DPPH, and FRAP results were 2.71 mg gallic acid equivalents (GAE)/g, 24 μmol Trolox equiv/g, and 22 μmol Trolox eq/g, respectively. The in vitro digestion process did not cause significant differences in TPC. However, the antioxidant activity was significantly decreased. Moreover, the bioavailability of anthocyanins was studied, showing that a small fraction of polyphenols in their intact form was conserved at the end of digestion. Finally, a protective effect of digested maize polyphenols was observed in the Caco-2 cell viability. The results suggest that "Moragro" purple maize is a good source of bioavailable anthocyanins in the diet and an interesting source of this group of compounds for the food industry.

Anti-melanogenic effect of <i>Amaranthus tricolorextract</i> on UV radiation-exposed melanoma cells

Purpose: To determine the inhibitory effect of A. tricolor extract (ATE) on melanogenesis in UV radiation-exposed B16F10 cells. 
 Methods: Total phenolic and anthocyanin contents of ATE were examined by Folin-Ciocalteu and pH differential methods, respectively. The tyrosinase inhibitory effect of ATE was investigated using L-DOPA as a substrate and the suppressive effect of ATE on melanin production was determined in melanoma cells. Furthermore, the antioxidant activity was assessed by DPPH method and reactive oxygen species formation while protein expression levels were evaluated by western blot method. 
 Results: The total phenolic and anthocyanin contents were identified up to 35 mg/g extract and 0.85 mg/g extract, respectively. ATE was found to inhibit both mushroom and cellular tyrosinase at IC50 values of 242.2 ± 9.5 μg/mL and 202.9 ± 11.6 μg/mL, respectively. Moreover, ATE treatment reduced melanin production up to (30 ± 5) % and suppressed p38 MAPK phosphorylation at the concentration of 400 μg/mL. In addition, ATE significantly scavenged DPPH radical with EC50 value of 189.2 ± 8.9 μg/mL and abolished ROS formation in the cells. 
 Conclusion: These results indicate that A. tricolor extract has a potential protective effect against melanin production, suggesting its potential value in skin-lightening formulations.

Phytochemical profiling and bioactivity study of Adenia panduriformis in Zambia using UHPLC-MS/MS-MZmine3, GNPS, and METLIN Gen2

Diseases, especially degenerative illnesses, can be treated and avoided altogether, and good health can be enhanced by including health-promoting foods in the diet without side effects. Adenia panduriformis (Passifloraceae) is a wild vegetable consumed as part of the everyday diet in some communities in Zambia. The plant is also used in traditional medicine to effectively treat and manage many diseases, including degenerative diseases. Because of significant health benefits and medicinal attributes, this vegetable has very high merit to be investigated for its phytochemicals and bioactivities. Therefore, this study screened phytochemicals using qualitative chemical tests and Liquid chromatography-tandem mass spectrometry (LCMS/MS), identified metabolites using molecular networking tools and evaluated the total polyphenolic content and antioxidant activity of the methanolic leaf extract of the vegetable. Solvent extraction by maceration recovered metabolites for screening by qualitative chemical tests, extraction by QuEChERS (quick, easy, cheap, effective, rugged, and safe) was used to recover metabolites for screening by LCMS/MS, and tentative phytochemical constituents were identified using MZmine3, GNPS (global natural products social molecular networking) and METLIN. Total phenolic content (TPC) determination used the Folin-Ciocalteu (FC) method at 765 nm. Total flavonoid content (TFC) was evaluated using the aluminium chloride colorimetric assay at 510 nm. Antioxidant activity was evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay at 517 nm. Phytochemical screening by qualitative chemical tests revealed the presence of many classes of phytocompounds, such as alkaloids, polyphenols, terpenoids, and saponins. TPC was determined to be 122.30 ± 2.00 mg GAE/g, and TFC was 60.17 ± 0.25 mg QE/g. Screening by LCMS/MS revealed the presence of many metabolites, of which 23 phytocompounds were identified using MZmine3, 42 by GNPS, and 21 by METLIN. The crude extract exhibited high DPPH free radical quenching activity with a relatively low IC50 value of 53.48 µg/mL compared to the reference standard ascorbic acid IC50 of 74.47 µg/mL. The significantly lower IC50 value for the wild vegetable than that of ascorbic acid indicates a considerably high efficiency of the metabolites in the vegetable as antioxidants. The presence of many metabolites with health-promoting properties and high antioxidant activity of the leaf extract of A. panduriformis may be the science behind its remarkable health benefits and power to treat and manage a variety of diseases, including degenerative illnesses. This study presents results that indicate the potential of A. panduriformis as a rich source of metabolites with brilliant health-promoting properties and high antioxidant activity, which may open exciting opportunities for further development in the fields of pharmacology and drug development.

RELATIONSHIP BETWEEN BIOCHEMICAL INDICATORS AND PHENOTYPICAL CHARACTERISTICS OF THYME (THYMUS VULGARIS L.)

Introduction. Thyme (Thymus vulgaris L.) is a sought-after medicinal crop, the raw materials of which are included in the Pharmacopoeias of several countries and are widely used in scientific medicine. Due to the presence of strong intraspecific diversity in thyme, it seems interesting to identify phenotypic characteristics that most likely indicate the presence of certain pharmacologically significant compounds. The presence of such information will allow targeted selection both in natural populations and during the hybridization of cultivars and the development of new varieties. The purpose of the work was to identify phenotypic traits indicating a high content of essential oil and thymol in thyme, the amount of phenolic compounds, flavonoids and tannins. Material and methods. Samples and varieties of thyme from different geographical origins were selected as objects. Biometric, anatomical indicators and yield were determined in the flowering phase. The essential oil content was determined by Clevenger hydrodistillation method. Component composition was determined by gas chromatography. The amount of phenolic compounds and tannins according to the Folin-Ciocalteu method in aqueous-alcoholic extract calculated as gallic acid. The determination of flavonoids in the aqueous-alcoholic extract was carried out using the spectrophotometryс method calculated as rutin. Results. As a result of the research, no signs were found indicating a high content of essential oil. Flavonoid content calculated as rutin had a negative correlation (−0.72) with stomatal density. A negative relationship was noted between the intensity of coloring of the pistil and anthers and the content of the total phenolic compounds calculated as gallic acid (0.71 and 0.72, respectively). Large stomata on both the underside and upper side of the leaf may indicate a high tannin content (0.8 and 0.88). All studied components of common thyme belonged to the thymol type, and the content of this component ranged from 37.6 to 80.8%. An increased content of thymol in essential oil is observed such signs as an elongated leaf shape (leaf index R = 0.78), lack of downward curling of the edges of the leaf blade (R = –0.78) and pronounced veins on the underside of the leaf (R = 0.92). Large stomata on the lower epidermis also indicate an increased proportion of thymol in the essential oil (R = 0.78). When analyzing the interdependence between the content of the main components of essential oil, it was noted that there is a close negative correlation between the content of p-cymene and thymol (R = –0.91) and carvacrol (R = –0.88). Conclusion. As a result of the research, phenotypic and anatomical characteristics were identified that make it possible to diagnose promising samples of common thyme, which will help speed up the selection process.

Physicochemical Properties of Apple Purees and Peel Extract for Potential Use in Pastry Products

In recent years, the consumption of pastry and bakery products has grown considerably, and consumers are increasingly tempted to choose products from an organoleptic point of view. At the same time, consumers are also interested in having a healthy diet, respectively, products with special sensory properties, but with a low sucrose content. Substituting the sucrose in these products with apple puree represents an alternative to obtaining cakes with a lower sugar content and, in addition, in obtaining products with high nutritional value, with the bioactive compounds from apples having special properties on health. The purpose of this work was to analyze both the physicochemical properties, the total content of polyphenols and the antioxidant activity of the apple puree samples, as well as their variation during the storage period. The physicochemical properties analyzed were: moisture content, titratable acidity, ash content, pH, water activity, total soluble solids content and color. Regarding the content of the bioactive compounds, the total content of polyphenols and the antioxidant capacity were determined by the Folin–Ciocalteu method, respectively, the DPPH method. The results showed that apple puree is an important source of polyphenols, and these are the main factors influencing antioxidant activity. The analysis of the properties of the three products obtained from the apple will allow you to choose one of them or a combination of them in order to obtain the highest degree of sugar substitution and the highest nutritional value of the products. The degree of substitution will be correlated with technological parameters, baking temperature and time.

Isolation of Phenolic Acids from Land Kale (Ipomoea reptans Poir) and Antioxidant Activity

Land kale or Ipomoea reptans Poir is widely consumed by Indonesian people. Land kale plants can be used as natural antioxidants because they contain polyphenolic compounds, one of which is phenolic acid. This research was carried out to determine the antioxidant activity and isolate phenolic acid compounds contained in land kale plants (Ipomoea reptans Poir). The sample used was an ethanol extract of land kale. The total phenolics were determined using the Folin-Ciocalteu method. Phenolic acids were isolated using alkaline hydrolysis and acid hydrolysis and without hydrolysis methods. Separation of isolates was carried out using the TLC method. The structure was identified using UV-Vis spectrophotometry and LC-MS/MS. Antioxidant activity was measured using the DPPH method. The total phenolics of land kale ethanol extract were 71.2420 ± 0.0791 mg/g GAE. TLC showed that the HB, HA, and TH fractions contained caffeic acid and ferulic acid compounds. Separation of the (HB) fraction produced three isolates: B2 with a yellow color, B3 with a slightly yellowish color, and B4 which is colorless. Isolate B2 was identified as potentially containing caffeic acid, while isolate B3 was indicated to potentially contain ferulic acid through analysis using a UV-Vis spectrophotometer. Isolate B4 was thought to have a hydroxybenzoic acid framework after being determined using LC-MS/MS. The ethanol extract of land kale has strong antioxidant activity because it produces an IC50 value of 94.83 mg/L.

Effects of Particle Size on Physicochemical and Nutritional Properties and Antioxidant Activity of Apple and Carrot Pomaces.

The food processing industry is growing rapidly and producing large amounts of by-products, such as pomaces, which are considered as no-value waste and cause significant environmental pollution. The main by-products of fruit juice processing companies are apple and carrot pomaces, which can be used to create new functional food products. In the present study, the effects of particle size (PS) on the proximate composition, nutritional properties, and antioxidant activity of apple pomace flour (APF) and carrot pomace flour (CPF) were determined. Four different PS fractions, PS > 1 mm, 1 > PS > 0.71 mm, 0.71 > PS > 0.18 mm, and 0.18 > PS > 0.075 mm were used for the present study. Their vitamin, carotenoid, organic acid, and reducing sugar contents were determined using HPLC. The proximate compositions of each PS fraction of the AP and CP flours were determined using recommended international standard methods. DPPH, FRAP, and Folin-Ciocalteu methods were used to measure their antioxidant activity and total phenolic compounds, respectively. The moisture content (around 12.1 mg/100 g) was similar in all PS fractions and in both flours. The APF had lower protein (4.3-4.6 g/100 g dw) and ash (1.7-2.0 g/100 g dw) contents compared to the CPF, with protein contents ranging from 6.4-6.8 g/100 g dw and ash contents ranging from 5.8-6.1 g/100 g dw. Smaller particles, regardless of flour type, exhibited higher sugar and phenolic contents and antioxidant activity, while vitamins were more abundant in particles larger than 1 mm. In the APF, larger particles had a higher fiber content than smaller particles, while their fat content was the lowest. PS also had an impact on the results of the carotenoid contents. This study underscores the direct impact of PS on the distribution of sugars, crude fiber, fat, carotenoids, vitamins, total phenolic compounds, and antioxidant activity in pomaces.

Modern and traditional cooking methods affect the antioxidant activity and phenolic compounds content of Trachystemon Orientalis (L.) G. Don.

Trachystemon orientalis (L.) G. Don is a medicinal plant with beneficial effects on human health. Its antioxidant and phenolic compound content is higher than most natural plants. This is the first study on the cooking of this consumed plant. This study investigated how different cooking methods and times affect the antioxidant activity and phenolic compound content of Trachystemon orientalis (L.) G. Don. The Folin-Ciocalteu method (FCR), ferric-reducing antioxidant power (FRAP), copper-reducing antioxidant capacity (CUPRAC), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity were used to evaluate the antioxidant activity and total phenolic content (TPC). Phenolic compounds were also determined by high-performance liquid chromatography (HPLC). Microwave cooking, stir-frying and sous vide increased TPC and antioxidant activity (p<0.05). Steaming decreased TPC and antioxidant activity (p<0.05). It was determined that the best cooking method and time was stir-frying for 15 minutes (TPC, CUPRAC and FRAP values 45.18±3.91 mg GAE/g DW, 15559.39±106.90 mmol Troloks/g DW and 555.10±24.05 μmol Fe (II)/g DW, respectively). Raw Trachystemon orientalis (L.) G. Don was detected with caffeic acid (31.53±0.25 mg/100 g DW). New phenolic compounds (protocatechuic acid and p-coumaric acid) were formed by boiling, stir-frying, microwaving, and sous vide methods. In conclusion, regarding antioxidant activity and phenolic compounds of Trachystemon orientalis (L.) G. Don; the best cooking methods are microwave, stir-frying, and sous vide (p<0.05). The most wrong cooking method is steaming (p<0.05).

Effectiveness of ammonium-based deep eutectic solvents in extracting polyphenol from Chlorella vulgaris

This study investigated the extraction of polyphenols from Chlorella vulgaris microalgae using environmentally friendly choline chloride-based deep eutectic solvents (DESs), specifically DES 1 (choline chloride: ethylene glycol, 1:2) and DES 2 (choline chloride: glycerol, 1:2). Thermal characterization of DESs through DSC analysis confirmed their formation as a glass former, while through density and viscosity analysis reveals their influence on extraction efficiency. Both DESs and methanol were employed in solid-liquid extraction combined with centrifugation. The Folin-Ciocalteu method quantified total phenolic content, showing DES 1 and DES 2 to outperform methanol (80.04, 68.63, and 60.15 mg GAE/g, respectively). High performance liquid chromatography (HPLC) was used in order to confirm these results. The HPLC result revealed that DES 1 exhibited the highest extraction efficiency at 198.00 ± 4.45 mg/L of Gallic acid yielded, followed by DES 2 at 189.83 ± 4.84 mg/L and the lowest is MeOH at 138.47 ± 5.36 mg/L. DPPH Radical Scavenging assay indicated both DES 1 and DES 2 extracts with significant radical scavenging activity (88.45 ± 0.17) % and (66.13 ± 1.62) %, respectively, compared to MeOH extracts that showed the lowest antioxidant activity at (30.41 ± 1.70) %. Overall, this study highlighted the potential of DESs in enchancing polyphenol extraction from Chlorella vulgaris, with room for further optimization and exploration of DES-polyphenol interactions.

Total phenolic and flavonoids content, and antioxidant activity of kratom (Mitragyna speciosa Korth.) leaf ethanol extract

This study aims to determine and compare antioxidant activity, total phenolics, flavonoids, and the relationship of phenolic and total flavonoids with the antioxidant activity of the three kratom variants. The ethanol extract of the third variant of kratom leaves was obtained by maceration. Through the antioxidant activity test using DPPH method, all three extracts showed strong antioxidant activity with IC50 values of 26.39, 30.25, and 30.59 μg/mL. The relationship of total phenolics and flavonoids with antioxidant activity was determined by the Pearson correlation test. Examination of total phenolic content using the Folin-Ciocalteu method with successive results for green, red, and white kratom are 6.11, 8.67, and 9.09 mg GAE/g extract. Examination of total flavonoid content using the colorimetric method for green, red, and white kratom were 0.86, 0.68, and 1.13 mg QE/g extract, respectively. The total phenolic content and antioxidant activity showed a correlation coefficient value of -0.32 (P0.05), and the total flavonoid content to antioxidant activity showed a correlation coefficient value of 0.81 (P0.05). Thus, all three variants of kratom have potency as natural antioxidants, but their total phenolic and flavonoid content does not influence their antioxidant activity.

Extraction of macroalgae phenolic compounds for cosmetic application using eutectic solvents

Cosmetic industry remains a major focus of economic development in the 21st century, while consumer's demand for sustainability drives the search for new active natural ingredients and the development of green extractions techniques. The aim of this work is to obtain antioxidant-enriched extracts from two seaweeds found on the Portuguese shore, Sargassum muticum and Gelidium corneum using natural deep eutectic solvents (NADES) to be used in cosmetic formulations. Extracts were obtained by maceration of powdered seaweeds with aqueous solutions (25 % (v/v) in water) of different NADES, L-lactic acid: fructose (5:1), L-lactic acid: glucose (5:1) and L-lactic acid: sodium acetate (7:1). The resulting extracts were evaluated regarding their total phenolic content (TPC) using the Folin-Ciocalteu method and antioxidant capacity by the DPPH and FRAP assays. L-lactic acid: fructose (5:1) NADES yielded Sargassum muticum extracts with the highest phenolic content (2099 mg GAE/L) and the highest antioxidant activities (DPPH reduction of 27 %, FRAP 30 μM FeSO4). Antimicrobial activity of the extracts over three microorganisms of the skin microbiota, the bacteria Staphylococcus epidermidis and Cutibacterium acnes and the fungus Malassezia furfur, revealed no relevant antimicrobial effect, meaning that extracts did not affect the skin microbiota homeostasis. The extract with the highest cosmeceutical interest, S. muticum extract prepared using L-lactic acid: fructose (7:1), was incorporated into a topical O/W emulsion. Physical and rheology data of the cosmetic formulation ensure its quality and safety. Finally, compatibility, acceptability and antioxidant efficacy were evaluated in vivo, in human volunteers. The results show that NADES are a promising replacement of conventional solvents in the extraction of antioxidants from biomass and their use in topical formulations is safe.

GAS CHROMATOGRAPHY-MASS SPECTROMETRY CHEMICAL PROFILING AND ANTIOXIDATION ACTIVITY OF THE THAI HERBAL FORMULA “PIKAD TRISUKKHATI SAMUTHARN” EXTRACT

Pikad Trisukkhati Samutharn (PTS) is a medicinal formula used in Thai traditional medicine for treating severe fever, inflammation, and diarrhea, as well as for general health promotion. This herbal formula was made from the roots of three plants: Oroxylum indicum (L.) Kurz, Sesbania grandiflora (L.) Pers and Ficus racemosa L. The purpose of this study was to investigate the antioxidation activity and chemical profiling of PTS and its herbal components. The antioxidation activity of the extracts was determined using the DPPH free radical scavenging assay, as well as the Folin-Ciocalteu method, to investigate the total phenolic contents. The thiobarbituric acid reactive substance (TBAR) assay was used to test lipid peroxidation activity. The chemical profiling of PTS was demonstrated using gas chromatography–mass spectrometry (GC-MS) analysis. The results revealed that the total phenolic content of the ethanolic extract (79.57 ± 2.33 mg GAE/g extract) was greater than that of the aqueous extract (12.55 ± 2.83 mg GAE/g extract) of the PTS. The DPPH free radical scavenging activity of the PTS ethanolic extract (IC50 value 80.69 ± 0.40 mg/mL) was also stronger than that of the PTS aqueous extract (IC50 value 650.10 ± 98.20 mg/mL). Lipid peroxidation of the PTS ethanolic extract was moderate with 32.47 ± 0.71% inhibition. GC-MS revealed the presence of several phenolic compounds contained in the PTS ethanolic extract in which the identified compounds had previously been reported to have several pharmacological activities such as antioxidation, anti-inflammation, and antimicrobial. In conclusion, the antioxidation and chemical constituents of the PTS extract were demonstrated for the first time, and more pharmacological activities of PTS should be explored to confirm its traditional use.

Antioxidant and color properties of black soybean tempe beverage with the addition of sappan wood and cinnamon

Black soybean was fermented into Tempe to increase its functional properties and bioavailability. To develop black soybean tempe into a beverage for practical consumption, sappan wood and cinnamon were added to improve the appearance and functional properties. This research aims to determine the effect of sappan wood and cinnamon addition on antioxidant levels, phenolic levels, and brightness levels of black soybean tempe beverages. The research used a Completely Randomized Design (CRD) with a ratio of sappan wood and cinnamon at 0:0, 3:0.5, 2.5:1, and 2:1.5 (%w/w). Beverage made from commercial yellow soybean tempe was used as control. This research started by making tempe using Yogyakarta's traditional method. Beans were dehulled, boiled for 30 minutes, soaked in boiling water for 36 hours, boiled again for 30 minutes, inoculated with RAPRIMA, and incubated for 40 hours at room temperature. Tempe beverage was made by milling tempe and 80°C water at 1:3 (w/w) and the barks at the mentioned ratio, followed by filtration. The beverages were then analyzed for their Ascorbic Acid Equivalent Antioxidant Capacity (AEAC), total phenolics using the Folin-Ciocalteu method, and brightness levels using the CIE method. The results showed the antioxidant properties ranged between 82.67-86.93 mg AEAC/ml, total phenolics: 53.43-66.53 mg GAE/ml, color brightness parameter L*: 33.82-34.49, a *: 1.97-2.01, and b*: 4.09-5.05. It can be concluded that adding sappan wood and cinnamon improved the antioxidant levels and appearance of black soybean tempe beverages. The highest antioxidant properties and brightness level were obtained by adding 3% and 0.5% of sappan wood and cinnamon bark, respectively.

Formulation and Evaluation of Herbal Antioxidant Tablets of Urtica Dioica Extract

This study explores the formulation and assessment of herbal antioxidant tablets derived from Urtica dioica (stinging nettle) extracts for arthritis and osteoarthritis treatment. Various extraction methods were compared, revealing that 100% dehydrated ethanol at 45 degrees Celsius for four hours in a Soxhlet apparatus resulted in an 89% phenolic extraction. Although ethyl acetate yielded the highest phenolic extract (187%), its toxicity led to the selection of ethanol for further extraction. The Folin-Ciocalteu method determined the total phenolic content. The DPPH assay demonstrated positive antioxidant activity of the Urtica dioica ethanolic extract, with an IC50 value of 37.5761201 μg/mL, compared to the standard ascorbic acid with an IC50 value of 21.23506 μg/ml. Herbal tablets were formulated using the direct compression method. The findings highlight the potential of Urtica dioica extracts as effective herbal antioxidants for arthritis and osteoarthritis treatment. This research enhances our understanding of natural remedies and their therapeutic applications. Future investigations should delve into the clinical efficacy and safety of these herbal antioxidant tablets, paving the way for their potential integration into clinical settings. This study contributes valuable insights into the development of alternative therapies for managing arthritic conditions, emphasizing the importance of exploring nature's remedies in healthcare.

Assessment of antioxidant potential, cytotoxicity, and anticancer activity of methanolic extracts from selected wild medicinal plants

IntroductionMedicinal plants are considered an important source of human health because of their therapeutic capabilities in treating various diseases. This study aimed to evaluate the phytochemical profile, antioxidant, and cytotoxic activities of the methanolic extracts of three wild medicinal plants (Ruta graveolens, Peganum harmala and Citrullus colocynthis). MethodsThe total phenolic content was determined according to the Folin–Ciocalteu method. The total flavonoid contents were measured by the aluminum chloride colorimetric method. The antioxidant activity was estimated based on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and ferric reducing antioxidant power (FRAP) assays. The phytochemical analysis was performed using liquid chromatography-mass spectrometry method. The cytotoxicity and anticancer activity of each plant extract was investigated against fibroblast and MDA-MB-231 breast cancer cell lines, respectively, using Giemsa staining assay. ResultsR. graveolens extract exhibited the highest concentrations of total phenols (25.5 ± 0.5 mg GA/g dry extract) and flavonoids (9.8 ± 0.5 mg Rutin/g dry extract). Additionally, it demonstrated the most robust antioxidant capacity, as demonstrated by both DPPH (100.8 ± 0.56 mg GA/g dry extract) and FRAP (91.2 ± 1.33 mg GA/g dry extract) assays. Within this plant, Rutin constituted 13.7 % of the total compounds. Furthermore, R. graveolens displayed the most significant anticancer activity, with statistical significance at p< 0.01. Notably, it exhibited the lowest cytotoxicity against the fibroblast cell line, even at the highest tested concentration of 100 μg/ml. In contrast, both P. harmala and C. colocynthis demonstrated substantial anticancer potential (p< 0.01); however, they both exhibited significant cytotoxicity against the fibroblast cell line (p<0.01). ConclusionR. graveolens extract holds promise as a potential reservoir of safe and effective antioxidant and anticancer compounds for human use.

Antioxidant and Antifungal Activities and Characterization of Phenolic Compounds Using High-Performance Liquid Chromatography and Mass Spectrometry (HPLC-MS) in Empetrum rubrum Vahl ex Willd.

In searching for compounds with antioxidant and antifungal activity, our study focused on the subshrub species Empetrum rubrum Vahl ex Willd. (Ericaceae). We measured the antioxidant activity of its methanolic extract (MEE) obtained from the aerial parts (leaves and stems) and of its methanolic extract (MEF) obtained from the lyophilized fruits. The antioxidant activity of the MEE and MEF was evaluated in vitro via a 2,2-Diphenyl-1-picrylhydrazyl (DPPH) free radical and 2,2'-Azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) cationic radical. The results were expressed in gallic acid and Trolox equivalents for the DPPH and ABTS assays, respectively. The antioxidant activities, for the DPPH and ABTS assays, were also evaluated by considering the IC50 values. Concerning the antioxidant activity, the total phenolic content (TPC) in the MEE and MEF was determined using the Folin-Ciocalteu method. Polyphenols contained in the leaves, stems, and fruits of E. rubrum were determined qualitatively by employing high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) analysis. The antifungal activity of the MEE obtained from the aerial parts of E. rubrum was tested against Rhizoctonia solani. The results of IC50 values measured by the DPPH and ABTS methods with MEE were 0.4145 ± 0.0068 mg mL-1 and 0.1088 ± 0.0023 mg mL-1, respectively, and the IC50 values for MEF were 6.4768 ± 0.0218 mg mL-1 and 0.7666 ± 0.0089 mg mL-1 measured by the DPPH and ABTS methods, respectively. The HPLC-MS analysis revealed the presence of anthocyanins, phenolic acids derivatives, and flavonols. In vitro, mycelial growth of this fungus was reduced from 90% to nearly 100% in the presence of MEE. The observed antifungal effect is related to the presence of the abovementioned phenols, detected in the MEE.

The Phenolic Content of Pistacia lentiscus Leaf Extract and Its Antioxidant and Antidiabetic Properties.

The aims of this study were to determine the polyphenolic profile, to estimate the total phenolic and flavonoid contents, and to evaluate the antioxidant and antidiabetic activities of the extract of Pistacia lentiscus leaves, and the hydroacetonic mixture was employed as an alternative for common solvents in the extraction process. In order to explain the antidiabetic activity, molecular docking has been performed on the main constituents of the leaf extract. The characterization of the extract has been performed by high-performance liquid chromatography (HPLC) leading to the detection of 20 compounds of which gallic acid, ellagic acid, catechin, kaempferol, and quercetin 3-glucoside were identified using authentic standards. The total phenolic and flavonoid contents, assessed using the Folin-Ciocalteu and quercetin methods, were 394.5 ± 0.08 mg gallic acid equivalent/g dry extract (mg GAE/g DE) and 101.2 ± 0.095 mg quercetin equivalent/g dry extract (mg QE/g DE), respectively. On the other hand, the antioxidant activity of leaf extract, quantified by determining the ability to neutralize the free radical DPPH and β-carotene/linoleate model system, reached the values of 0.0027 ± 0.002 mg/mL and 0.128 ± 0.04 mg/mL, respectively. Regarding the antidiabetic activity, based on the inhibition of pancreatic α-amylase activity, a significant inhibition of about 68.20% with an IC50 value of 0.266 mg/mL had been observed. This finding is consistent with the molecular docking study of the main phenolic compounds of the extracts, where a remarkable binding affinity against α-amylase was observed, with values of -7.631 (kcal/mol), -6.818 (kcal/mol), and -5.517 (kcal/mol) for the major compounds catechin, quercetin-3-glucoside, and gallic acid, respectively.

The Effects of Different Solvents on Phenolic, Flavonoid, Anthocyanin Contents and Free Radical Scavenging Activity on Pomegranate Juice and Seeds

Our phytochemical investigation defined the polyphenol, anthocyanin and flavonoid content from methanolic, ethanolic 50% and watery extracts from fresh pomegranate juice and seeds with a spectrophotometric method. The antioxidant capacity was defined with the ABTS and DPPH methods. We determined the polyphenols with the Folin-Ciocalteu method, the anthocyanins with the method in Hungarian Pharmacopoeia VIIIth edition at Myrtilli fructus monography and the flavonoids with a modified method of the Romanian Pharmacopeia Xth edition Cynarae folium monography. We expressed the concentration of polyphenols in gallic acids (GA). The values indicated the following: in fruit juice, 47.57 μg GA/ml, in methanol seed extract 7 μg GA/g, the ethanol 50% and in watery seed extracts 8 μg GA/g. The definition of flavonoids indicated the following values expressed in quercetin (QE): in fruit juice 706 μg QE/ml, in methanol seed extract 416 μg QE/g, in ethanol 50% seed extract 46 μg QE/g and in watery seed extract 57 μg QE/g. The anthocyans from juice 3.95 mg/100 ml and seed 6.47 mg/100 g expressed in cyanidin-3-0-glycoside. We determined the following values with the ABTS method: the antioxidant capacity of juice is 25.40 μg/ml, in methanol seed extract 2.87 μg/ml, in ethanol seed extract 2.52 μg/ml, in watery 9.15 μg/ml. With the DPPH method, we obtained the following values: from juice 54.2 μg/ml, from methanolic extract 14.7 μg/ml, from ethanolic extract 10.58 μg/ml and watery extract 18.22 μg/ml. The received data shows the importance of pomegranate, which can be a new phytotherapeutic potential resource.

Phytochemical and In-vitro Antioxidant Activity of Guiera senegalensis (Combretaceae) Leaf Extracts

Guiera senegalensis is a plant of the Combretaceae family, found in West Africa and Central Africa. It has many properties such as antibacterial, antifungal, anticancer, antioxidants, which are the subject of this study. The objective of this study was to realize a qualitative and quantitative phytochemical screening, evaluation of the antioxidant activity of the plant leaf extracts. Plant extractions was done in accordance with the methods described by Fonmboh and collaborators on the leaves of the plant; a qualitative phytochemical screening according to the methods described by Shaik and al quantitative phytochemical screening consisting of colorimetric dosage and finally the antioxidant activity was explored in-vitro by the Folin-Ciocalteu method and by the phosphomolybdenum method. This study revealed the presence of primary metabolites such as carbohydrates and proteins but also secondary metabolites such as alkaloids, flavonoids and coumarins. Quantification analysis demonstrated a significant concentration of these metabolites depending on the extract. Carbohydrates, proteins and polyphenols had a higher concentration in the hydro-ehtanolic extract with concentrations of 45,38 ± 1,88, 1172,31 ± 17,59, 310,27 ± 11,83 µg/ml respectively. Flavonoids, flavonols and alkaloids in the Aqueous maceration extract with concentrations of 145,84 ± 8,34, 169,52 ± 10,13, 320,56 ± 67,52 µg/ml. and Tannins in decoction extract with a concentration of 338,55 ± 12,25 µg/ml. The in-vitro antioxidant activity demonstrated an inhibition percentage of the order of 70.96 ± 0.58% in the extract obtained by hydro-ethanolic maceration. This study confirms the use of the plant in category 1 traditional medicine, and results show a promising potential for developing a category 2 phytomedicine. Future studies should explore the use of phytochemical compounds involved in this antioxidant activity as well as its mechanism.

Black Garlic (Allium sativum L.) Ethanol Extract: Total Phenolic Content and Antioxidant Activity Test Determination Using the Soxhletation Method

A plant that is commonly used in the food and health industries is garlic (Allium sativum L.). One powerful antioxidant found in garlic is called allicin. The garlic used in this study was heated to 70°C in order to produce black garlic. Determining the phenolic content overall and the antioxidant activity of the Black Garlic ethanol extract are the goals of this investigation. The Folin Ciocalteu method was used to calculate the total phenolic content, the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method was used to test for antioxidant activity, and the soxhletation method was used to extract black garlic. The findings indicated that the ethanol extract of black garlic had an overall phenolic content of 8.96% and an IC50 value of 4.7527 µg/mL (extremely strong antioxidant) for its antioxidant activity.